Methods for transduction and cell processing

A cell, axis-of-rotation technology, applied in the field of cell processing, capable of solving problems where available methods are not fully satisfactory

Active Publication Date: 2017-08-29
JUNO THERAPEUTICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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  • Methods for transduction and cell processing
  • Methods for transduction and cell processing
  • Methods for transduction and cell processing

Examples

Experimental program
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Effect test

Embodiment 1

[0602] Example 1: Viral transduction of primary human T cells in a centrifuge chamber

[0603] This example demonstrates the transduction of isolated primary human T cells with a recombinant viral vector encoding a chimeric antigen receptor (CAR), wherein, according to the embodiments provided herein, the transduction takes place in a substantially stationary cylindrical centrifuge chamber. Start under centrifugal force. T cells were isolated from human apheresis samples by positive selection.

[0604] The resulting cells were activated using anti-CD3 / CD28 reagents. To initiate transduction, cells were incubated after activation with viral particles containing the viral vector genome encoding the anti-CD19 CAR under various conditions.

[0605] Under a set of conditions ("Sepax"), by Transduction was initiated by incubating cells centrifuged in the cavity of a centrifuge chamber (Biosafe SA, A200) in a 2-processing unit (Biosafe SA). 50mL contains 50×10 6 The liquid comp...

Embodiment 2

[0608] Example 2: Transduction of primary human T cells in centrifuge chambers with different ratios of liquid volume to surface area

[0609] Transduction efficiency after initiation of transduction in the centrifuge chamber was assessed under various conditions using the same number of cells and virus infection units and different ratios of liquid volume to internal surface area of ​​the centrifuge chamber cavity. Cells were generally prepared and stimulated as described in Example 1. All transductions were initiated using a 2:1 IU:cell ratio with 100×10 6 total number of cells.

[0610] For the first sample ("5.1mL / sq.in." means 5.1:1mL liquid / square inch internal cavity surface used in this condition), containing 100×10 6 A liquid volume of 100 mL of cells is combined with 100 mL of the liquid composition containing the viral vector particles. For the second sample ("2.5 mL / in2" means 2.5 mL of liquid per square inch of cavity surface used in this condition), 50 mL of t...

Embodiment 3

[0612] Example 3: Transduction of primary human T cells in a centrifuge chamber

[0613] Another study compared transduction efficiency under various conditions, including transduction in centrifugation chambers using various ratios of liquid volume to cavity surface area, according to embodiments of the methods provided. Human T cells were isolated from apheresis products and stimulated as described above.

[0614] After stimulation, the 80×10 6 Individual cells were incubated under different conditions, including transduction with a CAR-encoding viral vector. Polycations were included in all samples.

[0615] Conditions for initiating transduction in the centrifuge chamber, 80 x 10 6 cells were incubated with vector-containing virus at a rate of 2 IU virus per cell in the cavity of a centrifuge chamber. currently using 2 The treatment system was incubated while centrifuging the centrifuge chamber for 60 minutes with an RCF of approximately 600 g on the inner side wall ...

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Abstract

Provided are methods, systems, and kits for cell processing, e.g., for therapeutic use, such as for adoptive cell therapy. The provided methods include transduction methods, in which cells and virus are incubated under conditions that result in transduction of the cells with a viral vector. The incubation in some embodiments is carried out in an internal cavity of a generally rigid centrifugal chamber, such as a cylindrical chamber made of hard plastic, the cavity of which may have a variable volume. The methods include other processing steps, including those carried out in such a chamber, including washing, selection, isolation, culture, and formulation. In particular, the disclosure relates to method providing advantages over available processing methods, such as available methods for large-scale processing. Such advantages include, for example, reduced cost, streamlining, increased efficacy, increased safety, and increased reproducibility among different subjects and conditions.

Description

[0001] Cross References to Related Applications [0002] This application claims U.S. Provisional Application No. 62 / 075,801, filed November 05, 2014, entitled "Methods for Transduction and Cell Processing," and the title filed March 05, 2015 Priority to US Provisional Application No. 62 / 129,023 for "Methods for Transduction and Cell Processing," the contents of which are incorporated by reference in their entirety. technical field [0003] The present invention relates to the treatment of cells for therapeutic use, eg for adoptive cell therapy. The provided methods generally include transduction methods in which cells and viral vector particles are incubated under conditions such that the cells are transduced with the viral vector. Incubation can be performed in the interior cavity of a generally rigid centrifuge chamber, such as a cylindrical centrifuge chamber made of hard plastic. The method includes other processing steps, including steps carried out in this centrifuge ...

Claims

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Application Information

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IPC IPC(8): C12N15/87C12N5/0783A61K35/17B04B7/08
CPCA61K35/17B04B7/08C07K14/7051C07K16/2896C12N5/0636C12N15/87C07K2319/74C12N15/86C12N2510/00A61K2300/00A61P31/00A61P35/00A61P37/06C07K2319/03A61K39/0011A61K2039/5156A61K2039/5158
Inventor R·L·克里斯曼C·拉姆斯伯格T·伍德
Owner JUNO THERAPEUTICS
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